Effects of Garcinia acid by down-regulation of Bcr-Abl protein on the proliferation and apoptosis of chronic myelogenous leukemia cell line K562
Huanliang Liu
Abstract
Huanliang Liu
Abstract
Objective To study the influence of Gambogic acid(GA) on the proliferation and cell apoptosis of chronic myeloid leukemia cells,and to investigate the possible mechanism.Methods After treatment with various concentrations of GA,cell viability was determined by MTS assay and trypan-blue counting method.The morphologic changes of K562 cells induced by GA were observed under fluorescence microscope with Propidium Iodide(PI) staining.Cell apoptosis was analyzed by Flow Cytometry staining with AnnexinV-FTIC / PI.The changes of apoptosis and proliferationrelated proteins were tested by Western blotting.Results GA inhibited the proliferation of K562 cells in a time-and dose-dependent manner(P 0.05).After treatment with various concentrations of GA,morphological changes were observed under fluorescence microscope,and nucleus of death cells were stained red with PI.Flow Cytometry showed that the death type of the treatment group was mainly apoptosis,and the apoptosis rate significantly increased compared with the control group(P 0.05).The caspase protein was activated,and Bcr-Abl protein and dow nstream signal pathway were inhibited(P 0.05).Conclusion GA could induce apoptosis and inhibit the proliferation of K562 cells.This effect may be mediated by activated caspase protein,reduced expression of Bcr-Abl protein and its dow nstream signal pathway.
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Objective To study the influence of Gambogic acid(GA) on the proliferation and cell apoptosis of chronic myeloid leukemia cells,and to investigate the possible mechanism.Methods After treatment with various concentrations of GA,cell viability was determined by MTS assay and trypan-blue counting method.The morphologic changes of K562 cells induced by GA were observed under fluorescence microscope with Propidium Iodide(PI) staining.Cell apoptosis was analyzed by Flow Cytometry staining with AnnexinV-FTIC / PI.The changes of apoptosis and proliferationrelated proteins were tested by Western blotting.Results GA inhibited the proliferation of K562 cells in a time-and dose-dependent manner(P 0.05).After treatment with various concentrations of GA,morphological changes were observed under fluorescence microscope,and nucleus of death cells were stained red with PI.Flow Cytometry showed that the death type of the treatment group was mainly apoptosis,and the apoptosis rate significantly increased compared with the control group(P 0.05).The caspase protein was activated,and Bcr-Abl protein and dow nstream signal pathway were inhibited(P 0.05).Conclusion GA could induce apoptosis and inhibit the proliferation of K562 cells.This effect may be mediated by activated caspase protein,reduced expression of Bcr-Abl protein and its dow nstream signal pathway.
Key concepts: Apoptosis, Propidium iodide, K562 cells, Flow cytometry, Molecular biology, Cell growth, Chronic myelogenous leukemia, Biology